<meta charset="utf8"></head><body>通过采血和激素分析评估狨猴的卵巢周期

<ol>
	<li>Kholkute, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasma+progesterone+levels+throughout+the+ovarian+cycle+of+the+common+marmoset+(Callithrix+jacchus).">Plasma progesterone levels throughout the ovarian cycle of the common marmoset (Callithrix jacchus).</a> Primates. 25 (1), 123-126 (1984).</li><li>Harding, R. D., Hulme, M. J., Lunn, S. F., Henderson, C., Aitken, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasma+progesterone+levels+throughout+the+ovarian+cycle+of+the+common+marmoset+(Callithrix+jacchus).">Plasma progesterone levels throughout the ovarian cycle of the common marmoset (Callithrix jacchus).</a> J Med Primatol. 11 (1), 43-51 (1982).</li><li>Gilchrist, R. B., Wicherek, M., Heistermann, M., Nayudu, P. L., Hodges, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+follicle-stimulating+hormone+and+follicle+populations+during+the+ovarian+cycle+of+the+common+marmoset.">Changes in follicle-stimulating hormone and follicle populations during the ovarian cycle of the common marmoset.</a> Biol Reprod. 64 (1), 127-135 (2001).</li><li>Gromoll, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+subclass+of+the+luteinizing+hormone/chorionic+gonadotropin+receptor+lacking+exon+10+messenger+RNA+in+the+New+World+monkey+(Platyrrhini)+lineage.">A new subclass of the luteinizing hormone/chorionic gonadotropin receptor lacking exon 10 messenger RNA in the New World monkey (Platyrrhini) lineage.</a> Biol Reprod. 69 (1), 75-80 (2003).</li><li>M&#252;ller, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chorionic+gonadotrophin+beta+subunit+mRNA+but+not+luteinising+hormone+beta+subunit+mRNA+is+expressed+in+the+pituitary+of+the+common+marmoset+(Callithrix+jacchus).">Chorionic gonadotrophin beta subunit mRNA but not luteinising hormone beta subunit mRNA is expressed in the pituitary of the common marmoset (Callithrix jacchus).</a> J Mol Endocrinol. 32 (1), 115-128 (2004).</li><li>Pacchiarotti, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovarian+stimulation+protocol+in+IVF:+an+up-to-date+review+of+the+literature.">Ovarian stimulation protocol in IVF: an up-to-date review of the literature.</a> Curr Pharm Biotechnol. 17 (4), 303-315 (2016).</li><li>Ezcurra, D., Humaidan, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+luteinising+hormone+and+human+chorionic+gonadotropin+when+used+in+assisted+reproductive+technology.">A review of luteinising hormone and human chorionic gonadotropin when used in assisted reproductive technology.</a> Reprod Biol Endocrinol. 12, 95 (2014).</li><li>Lopez-Gatius, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovarian+response+to+prostaglandin+F(2alpha)+in+lactating+dairy+cows:+A+clinical+update.">Ovarian response to prostaglandin F(2alpha) in lactating dairy cows: A clinical update.</a> J Reprod Dev. 68 (2), 104-109 (2022).</li><li>Marini, R. P., Wachtman, L. M., Tardif, S. D., Mansfield, K., Fox, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Common Marmoset in Captivity and Biomedical Research. , (2018).</li><li>Schultz-Darken, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sample+collection+and+restraint+techniques+used+for+common+marmosets+(Callithrix+jacchus).">Sample collection and restraint techniques used for common marmosets (Callithrix jacchus).</a> Comp Med. 53 (4), 360-363 (2003).</li><li>Hopper, J., Kubik, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Common+marmosets.">Common marmosets.</a> Handbook of Exotic Pet. , 27-42 (2020).</li><li>Harlow, C. R., Hearn, J. P., Hodges, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovulation+in+the+marmoset+monkey:+endocrinology,+prediction+and+detection.">Ovulation in the marmoset monkey: endocrinology, prediction and detection.</a> J Endocrinol. 103 (1), 17-24 (1984).</li><li>Ludlage, E., Mansfield, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+care+and+diseases+of+the+common+marmoset+(Callithrix+jacchus).">Clinical care and diseases of the common marmoset (Callithrix jacchus).</a> Comp Med. 53 (4), 369-382 (2003).</li><li>Summers, P. M., Wennink, C. J., Hodges, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cloprostenol-induced+luteolysis+in+the+marmoset+monkey+(Callithrix+jacchus).">Cloprostenol-induced luteolysis in the marmoset monkey (Callithrix jacchus).</a> J Reprod Fertil. 73 (1), 133-138 (1985).</li><li>Takahashi, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Birth+of+healthy+offspring+following+ICSI+in+in+vitro-matured+common+marmoset+(Callithrix+jacchus)+oocytes.">Birth of healthy offspring following ICSI in in vitro-matured common marmoset (Callithrix jacchus) oocytes.</a> PLoS One. 9 (4), e95560 (2014).</li><li>Tomioka, I., Takahashi, T., Shimada, A., Yoshioka, K., Sasaki, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Birth+of+common+marmoset+(Callithrix+jacchus)+offspring+derived+from+in+vitro-matured+oocytes+in+chemically+defined+medium.">Birth of common marmoset (Callithrix jacchus) offspring derived from in vitro-matured oocytes in chemically defined medium.</a> Theriogenology. 78 (7), 1487-1493 (2012).</li><li>Abe, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+marmoset+genome+engineering+by+autologous+embryo+transfer+and+CRISPR/Cas9+technology.">Efficient marmoset genome engineering by autologous embryo transfer and CRISPR/Cas9 technology.</a> Sci Rep. 11 (1), 20234 (2021).</li><li>Summers, P. M., Shephard, A. M., Taylor, C. T., Hearn, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+cryopreservation+and+transfer+on+embryonic+development+in+the+common+marmoset+monkey,+Callithrix+jacchus.">The effects of cryopreservation and transfer on embryonic development in the common marmoset monkey, Callithrix jacchus.</a> J Reprod Fertil. 79 (1), 241-250 (1987).</li><li>Thomson, J. A., Kalishman, J., Hearn, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonsurgical+uterine+stage+preimplantation+embryo+collection+from+the+common+marmoset.">Nonsurgical uterine stage preimplantation embryo collection from the common marmoset.</a> J Med Primatol. 23 (6), 333-336 (1994).</li><li>Hanazawa, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimally+invasive+transabdominal+collection+of+preimplantation+embryos+from+the+common+marmoset+monkey+(Callithrix+jacchus).">Minimally invasive transabdominal collection of preimplantation embryos from the common marmoset monkey (Callithrix jacchus).</a> Theriogenology. 78 (4), 811-816 (2012).</li><li>Ishibashi, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+embryo+transfer+in+the+common+marmoset+monkey+(Callithrix+jacchus)+with+a+reduced+transfer+volume:+a+non-surgical+approach+with+cryopreserved+late-stage+embryos.">Efficient embryo transfer in the common marmoset monkey (Callithrix jacchus) with a reduced transfer volume: a non-surgical approach with cryopreserved late-stage embryos.</a> Biol Reprod. 88 (5), 115 (2013).</li><li>Kishimoto, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+novel+common+marmoset+embryonic+stem+cell+lines+under+various+conditions.">Establishment of novel common marmoset embryonic stem cell lines under various conditions.</a> Stem Cell Res. 53, 102252 (2021).</li><li>Diehl, K. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+good+practice+guide+to+the+administration+of+substances+and+removal+of+blood,+including+routes+and+volumes.">A good practice guide to the administration of substances and removal of blood, including routes and volumes.</a> J Appl Toxicol. 21 (1), 15-23 (2001).</li><li>Daskalaki, M., Drummer, C., Behr, R., Heistermann, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+alfaxalone+for+short-term+anesthesia+can+confound+serum+progesterone+measurements+in+the+common+marmoset:+a+case+report.">The use of alfaxalone for short-term anesthesia can confound serum progesterone measurements in the common marmoset: a case report.</a> Primate Biol. 9 (2), 23-28 (2022).</li><li>Hodges, J. K., Cottingham, P. G., Summers, P. M., Liang, Y. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+ovulation+in+the+marmoset+monkey+(Callithrix+jacchus)+with+human+chorionic+gonadotropin+following+prostaglandin-induced+luteal+regression.">Controlled ovulation in the marmoset monkey (Callithrix jacchus) with human chorionic gonadotropin following prostaglandin-induced luteal regression.</a> Fertil Steril. 48 (2), 299-305 (1987).</li><li>Barrett, J., Abbott, D. H., George, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extension+of+reproductive+suppression+by+pheromonal+cues+in+subordinate+female+marmoset+monkeys,+Callithrix+jacchus.">Extension of reproductive suppression by pheromonal cues in subordinate female marmoset monkeys, Callithrix jacchus.</a> J Reprod Fertil. 90 (2), 411-418 (1990).</li><li>Barrett, J., Abbott, D. H., George, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensory+cues+and+the+suppression+of+reproduction+in+subordinate+female+marmoset+monkeys,+Callithrix+jacchus.">Sensory cues and the suppression of reproduction in subordinate female marmoset monkeys, Callithrix jacchus.</a> J Reprod Fertil. 97 (1), 301-310 (1993).</li></ol>

<table><tbody><tr><td>AIA-360</td><td>Tosoh Corporation</td><td>0019945</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>AIA-PACK DILUENT CONCENTRATE</td><td>Tosoh Corporation</td><td>0020956</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>AIA-PACK SUBSTRATE SET II</td><td>Tosoh Corporation</td><td>0020968</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>AIA-PACK WASH CONCENTRATE</td><td>Tosoh Corporation</td><td>0020955</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>CMS-1</td><td>CLEA Japan</td><td></td><td>Marmoset food</td></tr><tr><td>Estrumate</td><td>MSD Animal Health</td><td></td><td>PGF2alpha analog (cloprostenol)</td></tr><tr><td>Gonal-f Subcutaneous Injection 150</td><td>Merck Biopharma Co., Ltd.</td><td></td><td>FSH</td></tr><tr><td>Gonatropin for intramuscular injection 1000</td><td>ASKA Pharmaceutical Co., Ltd.</td><td>872413</td><td>hCG</td></tr><tr><td>Heparin sodium injection solution 5,000 units/5 mL</td><td>Mochida Pharmaceutical Co., Ltd.</td><td>224122458</td><td>Blood collection</td></tr><tr><td>Immunochromatographic Test Kit for Detection of Common Marmoset Chorionic Gonadotropin (Dual Checker)</td><td>CLEA Japan, Inc.</td><td></td><td>Determining CG level</td></tr><tr><td>Low-profile double-arm microscope illumination LPF-SD</td><td>SHIOKAZE GIKEN</td><td></td><td>Desk lamp for blood collection</td></tr><tr><td>Marmoset blood collection restraint device</td><td>JIC Japan</td><td>JM-1006</td><td>Blood collection&lt;br/&gt; http://www.jic-japan.jp/prd/marmoset/prd016.html&lt;br/&gt; email: vi@jic-japan.jp</td></tr><tr><td>Metacam 0.05%</td><td>Boehringer Ingelheim Animal Health Japan Co., Ltd.</td><td></td><td>Hematoma treatment</td></tr><tr><td>Sample Cup, 3 mL, PS, for Tosoh 360 and AIA-600 II, 1000/Bag</td><td>Globe Scientific</td><td>110913</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>ST AIA-PACK iE2</td><td>Tosoh Corporation</td><td>0025224</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>ST AIA-PACK iE2 CALIBRATOR SET</td><td>Tosoh Corporation</td><td>0025324</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>ST AIA-PACK iE2 SAMPLE DILUTING SOLUTION</td><td>Tosoh Corporation</td><td>0025524</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>ST AIA-PACK PROGIII</td><td>Tosoh Corporation</td><td>0025240</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>ST AIA-PACK PROGIII CALIBRATOR SET</td><td>Tosoh Corporation</td><td>0025340</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>ST AIA-PACK PROGIII SAMPLE DILUTING SOLUTION</td><td>Tosoh Corporation</td><td>&amp;nbsp;0025540</td><td>Hormone measurement (P4/E2)</td></tr><tr><td>Syringe with 25 G (0.50 x 25 mm) needle</td><td>TERUMO</td><td>SS-01T2525</td><td>Blood collection</td></tr><tr><td>Tensolvet 5.000 I.E. gel.</td><td>Dechra Pharmaceuticals</td><td>14033492</td><td>Hematoma treatment</td></tr><tr><td>TOSOH MULTI-CONTROL SET</td><td>Tosoh Corporation</td><td>0015965</td><td>Hormone measurement (P4/E2)</td></tr></tbody></table>

blood sampling and hormone measurement for determining the stage in the ovarian cycle in marmosets

<meta charset="utf8"></head><body>作者聚焦:通过生殖细胞研究和基因改造技术推进灵长类动物学

用于确定狨猴卵巢周期阶段的血液采样和激素测量

Common marmosets are small New World monkeys. Since many of their biological mechanisms are similar to those of humans, marmosets are potentially useful ...

blood-sampling-hormone-measurement-for-determining-stage-ovarian

Research

JoVE Journal

Biology

Blood Sampling and Hormone Measurement for Determining the Stage in the Ovarian Cycle in Marmosets

953 次观看.  National Center for Child Health and Development. 我们正在两个主要方面开展工作。首先，我们的目标是了解灵长类动物（包括人类）的雄性生殖细胞。我们专注于精原干细胞和 DNA 甲基化的建立。其次，我们的目标是开发制造转基因狨猴的新技术。转基因狨猴的产生需要付出很多努力。挑战之一是开发更有效的方法来制造转基因狨猴。我们的血液采样方案可以更轻松地找到静脉并降低从...

视频: 作者聚焦:通过生殖细胞研究和基因改造技术推进灵长类动物学

Protocol for Studying Extinction of Conditioned Fear in Naturally Cycling Female Rats

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that neural mechanisms for certain behavioral tasks and response behaviors are different in females and males. Using females in research studies can represent a challenge because of the variation of gonadal hormones during their estrous cycle. This protocol describes well-established procedures that are useful in investigating the role of estrogen in fear extinction memory consolidation in female rats. Phase of the estrous cycle and exogenous estrogen administration prior to extinction training can influence extinction recall 24 hr later. The vaginal swabbing technique for estrous phase identification described here aids the examination and manipulation of naturally cycling gonadal hormones. The use of this basic rodent model may further delineate the mechanisms by which estrogen can modulate fear extinction memory in females.

Gonadal hormones such as estrogen modulate memory formation in a number of experimental paradigms including fear extinction memory. This protocol describes a set of methods for investigating the influence of gonadal hormones specifically during extinction in naturally cycling females, including estrous cycle monitoring and exogenous estrogen administration.

协议在骑自行车自然雌性大鼠学习条件性恐惧灭绝

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that ...

科研

行为学

A Hyperandrogenic Mouse Model to Study Polycystic Ovary Syndrome

Hyperandrogenemia plays a critical role in reproductive and metabolic function in females and is the hallmark of polycystic ovary syndrome. Developing a lean PCOS-like mouse model that mimics women with PCOS is clinically meaningful. In this protocol, we describe such a model. By inserting a 4 mm length of DHT (dihydrotestosterone) crystal powder pellet (total length of pellet is 8 mm), and replacing it monthly, we are able to produce a PCOS-like mouse model with serum DHT levels 2 fold higher than mice not implanted with DHT (no-DHT). We observed reproductive and metabolic dysfunction without changing body weight and body composition. While exhibiting a high degree of infertility, a small subset of these PCOS-like female mice can get pregnant and their offspring show delayed puberty and increased testosterone as adults. This PCOS-like lean mouse model is a useful tool to study the pathophysiology of PCOS and the offspring from these PCOS-like dams.

We describe the development of a lean PCOS-like mouse model with&#160;dihydrotestosterone pellet to study the pathophysiology of PCOS and the offspring from these PCOS-like dams.

Hyperandrogenic 小鼠模型研究多囊卵巢综合征

Hyperandrogenemia plays a critical role in reproductive and metabolic function in females and is the hallmark of polycystic ovary syndrome. Developing a ...

发育生物学

Lung microRNA Profiling Across the Estrous Cycle in Ozone-exposed Mice

MicroRNA (miRNA) profiling has become of interest to researchers working in various research areas of biology and medicine. Current studies show a promising future of using miRNAs in the diagnosis and care of lung diseases. Here, we define a protocol for miRNA profiling to measure the relative abundance of a group of miRNAs predicted to regulate inflammatory genes in the lung tissue from of an ozone-induced airway inflammation mouse model. Because it has been shown that circulating sex hormone levels can affect the regulation of lung innate immunity in females, the purpose of this method is to describe an inflammatory miRNA profiling protocol in female mice, taking into consideration the estrous cycle stage of each animal at the time of ozone exposure. We also address applicable bioinformatics approaches to miRNA discovery and target identification methods using limma, an R/Bioconductor software, and functional analysis software to understand the biological context and pathways associated with differential miRNA expression.

Here we describe a method to assess lung expression of miRNAs that are predicted to regulate inflammatory genes using mice exposed to ozone or filtered air at different stages of the estrous cycle.

在暴露于臭氧的小鼠的弹性周期中的肺微 rna 分析

MicroRNA (miRNA) profiling has become of interest to researchers working in various research areas of biology and medicine. Current studies show a ...

免疫与感染

Frequent Tail-tip Blood Sampling in Mice for the Assessment of Pulsatile Luteinizing Hormone Secretion

In many endocrine systems, circulating factors or hormones are not released continuously, but are secreted as a discrete pulse in response to a releasing factor. Single-point sampling measures are inadequate to fully understand the biological significance of the secretory pattern of pulsatile hormones either under normal physiologic conditions or during conditions of dysregulation. Luteinizing hormone (LH) is synthesized by the anterior pituitary gonadotrope&#160;cells and secreted in a pulsatile pattern which requires frequent collection of blood samples for pulse assessment. This has not been possible in mice until recently, due to the development of a high-sensitivity LH assay and advancement in a technique for frequent low-volume sample collection, initially described by Steyn and colleagues.1 Here we describe a protocol for the frequent peripheral blood sample collection from mice with sufficient handling acclimatization to detect pulsatile secretion of LH. The current protocol details an expanded acclimatization period that allows assessment of robust and continuous pulses of LH over multiple hours. In this protocol, the tip of the tail is clipped&#160;and blood is collected from the tail using a hand-held pipette. For assessment of pulsatile LH in gonadectomized mice, serial samples are collected every 5-6 min for 90-180 min. Importantly, the collection of blood and measurement of robust pulses of LH can be accomplished in awake, freely behaving mice, given adequate handling acclimatization and effort to minimize environmental stressors. Sufficient acclimatization can be achieved within 4-5 weeks prior to blood collection. This protocol highlights advances in the methodology to ensure collection of whole blood samples for assessment of pulsatile LH secretion patterns over multiple hours in the mouse, a powerful animal model for neuroendocrine research.

Here we present a tail-tip blood sampling protocol for frequent sample collection in unrestrained mice. This method is useful for assessing patterns of pulsatile luteinizing hormone secretion and could be adapted for analysis of other circulating factors.

小鼠经常尾端采血对脉动性促黄体激素分泌的评价

In many endocrine systems, circulating factors or hormones are not released continuously, but are secreted as a discrete pulse in response to a releasing ...

神经科学

Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification

A rapid means of assessing reproductive status in rodents is useful not only in the study of reproductive dysfunction but is also required for the production of new mouse models of disease and investigations into the hormonal regulation of tissue degeneration (or regeneration) following pathological challenge. The murine reproductive (or estrous) cycle is divided into 4 stages: proestrus, estrus, metestrus, and diestrus. Defined fluctuations in circulating levels of the ovarian steroids 17-&beta;-estradiol and progesterone, the gonadotropins luteinizing and follicle stimulating hormones, and the luteotropic hormone prolactin signal transition through these reproductive stages. Changes in cell typology within the murine vaginal canal reflect these underlying endocrine events. Daily assessment of the relative ratio of nucleated epithelial cells, cornified squamous epithelial cells, and leukocytes present in vaginal smears can be used to identify murine estrous stages. The degree of invasiveness, however, employed in collecting these samples can alter reproductive status and elicit an inflammatory response that can confound cytological assessment of smears. Here, we describe a simple, non-invasive protocol that can be used to determine the stage of the estrous cycle of a female mouse without altering her reproductive cycle. We detail how to differentiate between the four stages of the estrous cycle by collection and analysis of predominant cell typology in vaginal smears and we show how these changes can be interpreted with respect to endocrine status.

Here, we describe how to identify the stage of the murine reproductive (proestrus, estrus, metestrus, or diestrus) by simple, non-invasive collection and cytological assessment of vaginal smear samples. We further describe how vaginal cytology reflects circulating hormonal levels underlying transition through the murine reproductive cycle.

进行阴道灌洗，结晶紫染色法和阴道细胞学评价小鼠发情周期分期鉴定

A rapid means of assessing reproductive status in rodents is useful not only in the study of reproductive dysfunction but is also required for the ...

生物学

DREADDs 技术和自动血液采样

Remote Neuronal Activation Coupled with Automated Blood Sampling to Induce and Measure Circulating Luteinizing Hormone in Mice

Circulating luteinizing hormone (LH) levels are an essential index of the functioning of the hypothalamic-pituitary control of reproduction. The role of numerous inputs and neuronal populations in the modulation of LH release is still unknown. Measuring changes in LH levels in mice is often a challenge since they are easily disrupted by environmental stress. Current techniques to measure LH release and pulsatility require long-term training for mice to adapt to manipulation stress, certain restraint, the presence of the investigator, and working on individual animals, reducing its usefulness for many research questions.
This paper presents a technique to remotely activate specific neuronal populations using Designer Receptor Exclusively Activated by Designer Drugs (DREADDs) technology coupled with automated sequential blood sampling in conscious, freely moving, and undisturbed mice. We first describe the stereotaxic surgery protocol to deliver adeno-associated virus (AAV) vectors expressing DREADDs to specific neuronal populations. Next, we describe the protocol for carotid artery and jugular vein cannulation and postsurgical connection to the CULEX automated blood sampling system. Finally, we describe the protocol for clozapine-N-oxide intravenous injection for remote neuronal activation and automated blood collection. This technique allows for programmed automated sampling every 5 min or longer for a given period, coupled with intravenous substance injection at a desired time point or duration. Overall, we found this technique to be a powerful approach for research on neuroendocrine control.

作者聚焦:自动输注和血液采样对清醒小鼠进行精确的激素分析

Luteinizing hormone (LH) pulsatility is a hallmark of reproductive function. We describe a protocol for remote activation of specific neuronal populations linked to serial automated blood collection. This technique allows timed hormonal modulation, multiplexing, and minimizing manipulation effects on LH levels in conscious freely moving, and undisturbed animals.

远程神经元激活与自动血液采样相结合，以诱导和测量小鼠体内循环黄体生成素

Circulating luteinizing hormone (LH) levels are an essential index of the functioning of the hypothalamic-pituitary control of reproduction. The role of ...

Reproductive Techniques for Ovarian Monitoring and Control in Amphibians

Ovarian control and monitoring in amphibians require a multi-faceted approach. There are several applications that can successfully induce reproductive behaviors and the acquisition of gametes and embryos for physiological or molecular research. Amphibians contribute to one-quarter to one-third of vertebrate research, and of interest in this context is their contribution to the scientific community&#39;s knowledge of reproductive processes and embryological development. However, most of this knowledge is derived from a small number of species. In recent times, the decimation of amphibians across the globe has required increasing intervention by conservationists. The captive recovery and assurance colonies that continue to emerge in response to the extinction risk make existing research and clinical applications invaluable to the survival and reproduction of amphibians held under human care. The success of any captive population is founded on its health and reproduction and the ability to develop viable offspring that carry forward the most diverse genetic representation of their species. For researchers and veterinarians, the ability to monitor and control ovarian development and health is, therefore, imperative. The focus of this article is to highlight the different assisted reproductive techniques that can be used to monitor and, where appropriate or necessary, control ovarian function in amphibians. Ideally, any reproductive and health issues should be reduced through proper captive husbandry, but, as with any animal, issues of health and reproductive pathologies are inevitable. Non-invasive techniques include behavioral assessments, visual inspection and palpation and morphometric measurements for the calculation of body condition indices and ultrasound. Invasive techniques include hormonal injections, blood sampling, and surgery. Ovarian control can be exercised in a number of ways depending on the application required and species of interest.

The study of amphibian biology provides valuable information on the reproductive, physiological, embryological and developmental processes that drive organisms of many taxonomic groups. Here, we present a comprehensive guide on different methodologies that can be used to study ovarian control and monitoring in amphibians.

两栖动物卵巢监测和控制的生殖技术

Ovarian control and monitoring in amphibians require a multi-faceted approach. There are several applications that can successfully induce reproductive ...

Bovine Ovarian Cortex Tissue Culture

Follicle development from the primordial to antral stage is a dynamic process within the ovarian cortex, which includes endocrine and paracrine factors from somatic cells and cumulus cell-oocyte communication. Little is known about the ovarian microenvironment and how the cytokines and steroids produced in the surrounding milieu affect follicle progression or arrest. In vitro culture of ovarian cortex enables follicles to develop in a normalized environment that remains supported by adjacent stroma. Our objective was to determine the effect of nutritional Stair-Step diet on the ovarian microenvironment (follicle development, steroid, and cytokine production) through in vitro culture of bovine ovarian cortex. To accomplish this, ovarian cortical pieces were removed from heifers undergoing two different nutritionally developed schemes prior to puberty: Control (traditional nutrition development) and Stair-Step (feeding and restriction during development) that were cut into approximately 0.5-1 mm3 pieces. These pieces were subsequently passed through a series of washes and positioned on a tissue culture insert that is set into a well containing Waymouth&#39;s culture medium. Ovarian cortex was cultured for 7 days with daily culture media changes. Histological sectioning was performed to determine follicle stage changes before and after the culture to determine effects of nutrition and impact of culture without additional treatment. Cortex culture medium was pooled over days to measure steroids, steroid metabolites, and cytokines. There were tendencies for increased steroid hormones in ovarian microenvironment that allowed for follicle progression in the Stair-Step versus Control ovarian cortex cultures. The ovarian cortex culture technique allows for a better understanding of the ovarian microenvironment, and how alterations in endocrine secretion may affect follicle progression and growth from both in vivo and in vitro treatments. This culture method may also prove beneficial for testing potential therapeutics that may improve follicle progression in women to promote fertility.

In vitro culture of bovine ovarian cortex and the effect of nutritional Stair-step diet on ovarian microenvironment is presented. Ovarian cortex pieces were cultured for seven days and steroids, cytokines, and follicle stages were evaluated. The Stair-Step diet treatment had increased steroidogenesis resulting in follicle progression in culture.

牛卵巢皮质组织培养

Follicle development from the primordial to antral stage is a dynamic process within the ovarian cortex, which includes endocrine and paracrine factors ...

用于确定狨猴卵巢周期阶段的血液采样和激素测量

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远程神经元激活与自动血液采样相结合，以诱导和测量小鼠体内循环黄体生成素

远程神经元激活与自动血液采样相结合，以诱导和测量小鼠体内循环黄体生成素

两栖动物卵巢监测和控制的生殖技术

牛卵巢皮质组织培养