Name: Yazar Spotlight: Anne Diyet Eksikliklerinin Uzun Vadeli Yavru Sağlığı Sonuçları Üzerindeki Etkisinin İncelenmesi
Uploaded: 2024-06-28T13:50:00
Description: Maternal nutrition during pregnancy and lactation plays an important role in the neurodevelopment of offspring. One-carbon (1C) metabolism, which centers ...

Bu protokol onaylandı ve tüm hayvan denekler Midwestern Üniversitesi Kurumsal Hayvan Bakımı ve Kullanımı Komitesi'nin yönergelerine uygun olarak kullanıldı (Protokol 2983). Bu çalışmada 2 aylık (vücut ağırlığı, ~ 20 g) dişi C57BL / 6J fareleri kullanıldı. Genel olarak, üreme çağındaki dişi fareler kullanılabilir10. Çöp etkilerinden kaçınılmasını sağlamak için, her diyet için birden fazla dişi kullanılması gerekir11. Çalışmada kul...

Maternal Folik Asit ve Kolin Eksikliklerinin Yavru Sağlığına Etkisi

<ol>
	<li>Van Gool, J. D., Hirche, H., Lax, H., De Schaepdrijver, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Folic+acid+and+primary+prevention+of+neural+tube+defects:+A+review.">Folic acid and primary prevention of neural tube defects: A review.</a> Reprod Toxicol. 80, 73-84 (2018).</li><li>Ali, S. A., Economides, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Folic+acid+supplementation.">Folic acid supplementation.</a> Curr Opin Obstet Gynecol. 12 (6), 507-512 (2000).</li><li>Osterhues, A., Ali, N. S., Michels, K. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+folic+acid+fortification+in+neural+tube+defects:+a+review.">The role of folic acid fortification in neural tube defects: a review.</a> Crit Rev Food Sci Nutr. 53 (11), 575966 (2013).</li><li>Hurley, L., 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=Maternal+dietary+deficiencies+in+folic+acid+and+choline+result+in+larger+damage+volume,+reduced+neuro-degeneration+and+-inflammation+and+changes+in+choline+metabolites+after+ischemic+stroke+in+middle-aged+offspring.">Maternal dietary deficiencies in folic acid and choline result in larger damage volume, reduced neuro-degeneration and -inflammation and changes in choline metabolites after ischemic stroke in middle-aged offspring.</a> Nutrients. 15 (7), 1556 (2023).</li><li>Clementson, M., 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=Maternal+dietary+deficiencies+in+folic+acid+or+choline+worsen+stroke+outcomes+in+adult+male+and+female+mouse+offspring.">Maternal dietary deficiencies in folic acid or choline worsen stroke outcomes in adult male and female mouse offspring.</a> Neural Regen R.es. 18 (11), 2443-2448 (2023).</li><li>Pull, 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=Impact+of+maternal+dietary+folic+acid+or+choline+dietary+deficiencies+on+vascular+function+in+young+and+middle-aged+female+mouse+offspring+after+ischemic+stroke.">Impact of maternal dietary folic acid or choline dietary deficiencies on vascular function in young and middle-aged female mouse offspring after ischemic stroke.</a> Am J Physiol Heart Circ Physiol. 325 (6), H1354-H1359 (2023).</li><li>Virdi, S., Jadavji, N. 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+impact+of+maternal+folates+on+brain+development+and+function+after+birth.">The impact of maternal folates on brain development and function after birth.</a> Metabolites. 12 (9), 8766 (2022).</li><li>Barker, D. 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=Fetal+nutrition+and+cardiovascular+disease+in+later+life.">Fetal nutrition and cardiovascular disease in later life.</a> Br Med Bull. 53 (1), 96-108 (1997).</li><li>Barker, D. 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=Maternal+nutrition,+fetal+nutrition,+and+disease+in+later+life.">Maternal nutrition, fetal nutrition, and disease in later life.</a> Nutrition. 13 (9), 807-813 (1997).</li><li>Jackson, S. 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=Does+age+matter?+The+impact+of+rodent+age+on+study+outcomes.">Does age matter? The impact of rodent age on study outcomes.</a> Lab Anim. 51 (2), 160-169 (2017).</li><li>Jim&#233;nez, J. A., Zylka, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+litter+effects+to+enhance+rigor+and+reproducibility+with+rodent+models+of+neurodevelopmental+disorders.">Controlling litter effects to enhance rigor and reproducibility with rodent models of neurodevelopmental disorders.</a> J Neurodev Disords. 13 (1), 2 (2021).</li><li>Breeding and Reproduction of Mice - All Other Pets. Merck Veterinary Manual Available from: <a target="_blank" href="https://www.merckvetmanual.com/all-other-pets/mice/breeding-and-reproduction-of-mice">https://www.merckvetmanual.com/all-other-pets/mice/breeding-and-reproduction-of-mice</a> (2024)</li><li>Jena, S., Chawla, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+anatomy+and+physiology+of+laboratory+mouse.">The anatomy and physiology of laboratory mouse.</a> Essentials of Laboratory Animal Science: Principles and Practices. , 159-185 (2021).</li><li>. The anatomy of the laboratory mouse Available from: <a target="_blank" href="https://www.informatics.jax.org/cookbook/imageindex.shtml">https://www.informatics.jax.org/cookbook/imageindex.shtml</a> (2023)</li><li>Mortimer, T., Welz, P. S., Benitah, S. A., Sassone-Corsi, P., Koronowski, K. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collecting+mouse+livers+for+transcriptome+analysis+of+daily+rhythms.">Collecting mouse livers for transcriptome analysis of daily rhythms.</a> STAR Protoc. 2 (2), 100539 (2021).</li><li>Aboghazleh, R., 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=Rodent+brain+extraction+and+dissection:+A+comprehensive+approach.">Rodent brain extraction and dissection: A comprehensive approach.</a> MethodsX. 12, 102516 (2024).</li><li>Parasuraman, S., Raveendran, R., Kesavan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+sample+collection+in+small+laboratory+animals.">Blood sample collection in small laboratory animals.</a> J Parmacol Pharmacother. 1 (2), 87-93 (2010).</li><li>Jadavji, N. M., Deng, L., Malysheva, O. V., Caudill, M. A., Rozen, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MTHFR+deficiency+or+reduced+intake+of+folate+or+choline+in+pregnant+mice+results+in+impaired+short-term+memory+and+increased+apoptosis+in+the+hippocampus+of+wild-type+offspring.">MTHFR deficiency or reduced intake of folate or choline in pregnant mice results in impaired short-term memory and increased apoptosis in the hippocampus of wild-type offspring.</a> Neuroscience. 300, 1-9 (2015).</li><li>Pickell, L., 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=High+intake+of+folic+acid+disrupts+embryonic+development+in+mice.">High intake of folic acid disrupts embryonic development in mice.</a> Birth defects research. Birth Defects Res A Clin Mol Teratol. 91 (1), 8-19 (2011).</li><li>Mikael, L. G., Deng, L., Paul, L., Selhub, J., Rozen, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Moderately+high+intake+of+folic+acid+has+a+negative+impact+on+mouse+embryonic+development.">Moderately high intake of folic acid has a negative impact on mouse embryonic development.</a> Birth Defects Res A Clin Mol Teratol. 97 (1), 47-52 (2013).</li><li>Mikael, L. G., Pancer, J., Wu, Q., Rozen, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disturbed+one-carbon+metabolism+causing+adverse+reproductive+outcomes+in+mice+is+associated+with+altered+expression+of+apolipoprotein+ai+and+inflammatory+mediators.">Disturbed one-carbon metabolism causing adverse reproductive outcomes in mice is associated with altered expression of apolipoprotein ai and inflammatory mediators.</a> J Nutr. 142 (3), 411-418 (2012).</li></ol>

Bu protokolün araştırma laboratuvarlarında uygulanması sırasında, araştırmacıların deneylere başlamadan önce göz önünde bulundurmaları gereken birkaç önemli not vardır. İlk olarak, diyet üretiminde, deneysel çalışmanın başlamasından önce diyetin üretilmesi için yeterli teslim süresinin verilmesi gerekir. Teslim süreleri şirketten şirkete değişebilir ve diyet sevkiyatının da hesaba katılması gerekir. Üreme çağındaki (~ 2-2.5 aylık) dişi fareler çiftleşmeden 4-6 hafta önce ...

Hamilelik ve emzirme döneminde annenin beslenmesi, yavru gelişiminde hayati bir rol oynar. Tek karbonlu (1C) metabolizma, beslenme sinyallerini biyosentez, redoks homeostazı ve epigenetik ile bütünleştiren önemli bir metabolik ağdır. Folik asit ve kolin, nöral tüpün kapanmasında önemli bir rol oynayan 1C'nin merkezi bileşenleridir. Kadınların doğurganlık yıllarında folik asit ve kolin takviyesi yapmaları yaygın olarak tavsiye edilir1. Folik asidin fetal gelişimdeki kritik rolü açıkça belirlenmiş olsa da, ilgili altta yatan mekanizmalar belirsizliğini korumaktadır. Çocuk doğuran kadınların hamile kalmadan önce folik asit almaları önerilir, çünkü nöral tüpün kapanması ilk trimester2 içinde gerçekleşir. Folik asit veya kolin gibi 1C'nin diğer bileşenlerinde eksik olan hamile anneler, nükleotid sentezi ve erken gebelik döneminin gerektirdiği diğer süreçlere olan yüksek talep nedeniyle nöral tüp defektli yavruların doğum insidansı daha yüksektir3.
1C metabolizması alımında yetersiz olan anne diyetlerinin yavru sağlığı sonuçları üzerindeki uzun vadeli etkisi tam olarak anlaşılamamıştır. Laboratuvarlarımızdan ve diğerlerinden yapılan araştırmalar, folik asit veya kolin eksikliği olan anne diyetlerinin, klinik öncesi modeller 4,5,6 kullanılarak iskemik inme sonrası yavru sağlığı sonuçlarını gerçekten etkilediğini göstermiştir. Maternal diyetin etkisi, daha küçük beyin hacimlerinin ve değişmiş metilasyon seviyelerinin tanımlandığı klinik çalışmalarda da gösterilmiştir7. Sağlık ve Hastalığın Gelişimsel Kökenleri (DOHaD) teorisi, prospektif kronik hastalıkların utero 8,9'da programlandığını ve anne diyetinin önemli bir rol oynadığını göstermektedir.
Bu protokol, anne beslenmesinin yavruların sağlık sonuçları üzerindeki etkilerini incelemek amacıyla, hamilelikten önce dişi farelerde 1C diyet tüketimini değiştirme yöntemini ana hatlarıyla belirtmek için tasarlanmıştır.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1.5 mL tubes</td>
			<td>Eppendorf</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6 Female Mice</td>
			<td>Charles River</td>
			<td>27</td>
			<td>Mice can be bred in house as well.</td>
		</tr>
		<tr>
			<td>Choline Deficient Diet</td>
			<td>Envigo</td>
			<td>TD.06119</td>
			<td></td>
		</tr>
		<tr>
			<td>Control Diet</td>
			<td>Envigo</td>
			<td>TD.190790</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA tubes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Folic Acid Deficient Diet</td>
			<td>Envigo</td>
			<td>TD.01546</td>
			<td></td>
		</tr>
		<tr>
			<td>GraphPad prism</td>
			<td></td>
			<td></td>
			<td>https://www.graphpad.com/features</td>
		</tr>
		<tr>
			<td>Housing for rodents</td>
			<td>Lab Products, LLC</td>
			<td>One CageVentilated Racks &#38; Cages</td>
			<td></td>
		</tr>
		<tr>
			<td>Liquid chromatography-tandem mass spectrometry (LC-MS/MS)</td>
			<td>SCIEX, Framingham, MA</td>
			<td>5500 QTrap</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultra-performance liquid chromatography (UPLC)</td>
			<td>Shimadzu, Columbia, MD, USA</td>
			<td>Nexera</td>
			<td></td>
		</tr>
	</tbody>
</table>

modifying levels of maternal dietary folic acid or choline to study the impact of deficiencies on offspring health outcomes

Hamilelik ve emzirme döneminde annenin beslenmesi, yavruların nörogelişiminde önemli bir rol oynar. Folik asit ve kolinin yanı sıra diğer B vitaminlerini merkeze alan tek karbonlu (1C) metabolizma, gelişmekte olan fetüsün nöral tüpünün kapanması sırasında anahtar bir rol oynar. Bununla birlikte, hamilelik sırasındaki bu maternal beslenme yetersizliklerinin doğumdan sonra yavru sağlığı sonuçları üzerindeki etkisi nispeten tanımlanmamıştır. Ayrıca, folik asit veya kolindeki maternal diyet eksiklikleri, yavrulardaki diğer sağlık sonuçlarını etkileyebilir - bu da bunu değerli bir model haline getirir. Bu protokol, diyet değişiklikleri yoluyla dişi farelerde 1C metabolizmasında bir eksikliğe neden olma prosedürünü ana hatlarıyla belirtmeyi amaçlamaktadır. Dişiler sütten kesimde, 2 aylık olana kadar, çiftleşmeden 4-6 hafta önce diyetlere yerleştirilir ve hamilelik ve emzirme boyunca diyette kalırlar. Bu dişilerden elde edilen yavrular sağlık sonuçları açısından değerlendirilebilir. Dişiler yavru üretmek için birden çok kez kullanılabilir ve 1C metabolit ölçümlerini ölçmek için dişilerden dokular toplanabilir. Bu protokol, yavru sağlığı sonuçlarını incelemek için folik asit veya kolindeki maternal diyet eksikliklerinin nasıl indükleneceğine dair bir genel bakış sağlar.

Maternal nutrition during pregnancy and lactation plays a vital role in offspring development. One-carbon (1C) metabolism is a key metabolic network that integrates nutritional signals with biosynthesis, redox homeostasis, and epigenetics. Folic acid and choline are central components of 1C that play an important role in the closure of the neural tube. It is widely recommended for women to supplement with folic acid and choline during their childbearing years1. While the critical role of folic acid in fetal development is clearly established, the underlying mechanisms involved remain unclear. Women of childbearing are recommended to take folic acid prior to becoming pregnant since the closure of the neural tube occurs within the first trimester2. Pregnant mothers who are deficient in folic acid or other components of 1C, like choline, have a higher incidence of birthing offspring with neural tube defects due to the high demand for nucleotide synthesis and other processes required by the early gestational period3.
The long-term impact of maternal diets deficient in 1C metabolism intake on offspring health outcomes is poorly understood. Research from our laboratories, as well as others, has demonstrated that maternal diets deficient in either folic acid or choline do indeed impact offspring health outcomes after ischemic stroke using preclinical models4,5,6. The impact of maternal diet has also been shown in clinical studies where smaller brain volumes and altered methylation levels have been described7. The Developmental Origins of Health and Disease (DOHaD) theory suggests that prospective chronic diseases are programmed in utero8,9, and maternal diet plays an important role.
This protocol is designed to outline the method for altering 1C dietary consumption in female mice before pregnancy, with the purpose of examining the effects of maternal nutrition on the health outcomes of the offspring.

Yazar Spotlight: Anne Diyet Eksikliklerinin Uzun Vadeli Yavru Sağlığı Sonuçları Üzerindeki Etkisinin İncelenmesi

Eksikliklerin yavru sağlığı sonuçları üzerindeki etkisini incelemek için maternal diyet folik asit veya kolin seviyelerinin değiştirilmesi

We are interested in understanding how maternal dietary deficiencies in folic acid or choline during pregnancy and lactation impact long-term offspring health outcomes. We know that folic acid is really important for the closure of the neural tube in utero, but my laboratory is interested in what roles it plays after birth. We are reading studies that are showing that maternal diet during pregnancy and lactation does play an important role in offspring health.Dietary changes include high fat, one carbon metabolism, et cetera. These changes are sometimes seen in several generations of offspring. We are using a variety of tools to understand the long-term effects on offspring that come from moms with dietary deficiencies in folate and choline.Our most recent study is looking at metabolite changes in stool samples of offspring. We use behavioral neuroscience, biochemical assays, molecular biology, and epigenetic tools. My research group is the first one to show that maternal dietary deficiencies in either folic acid or choline during pregnancy and lactation result in worse stroke outcome for female and male offspring.We are really interested in understanding the long-term role that maternal dietary deficiencies in either folic acid or choline have on offspring health outcomes. Folic acid is well known for its role in the closure of the neural tube in utero. The impact of deficiencies after birth on brain function are not well-defined.

Bu çalışmanın odak noktası, maternal diyet eksikliklerinin folik asit veya kolin üzerindeki yavru sağlığı sonuçları üzerindeki etkisini araştırmaktı. Folik asit veya kolin eksikliği olan diyetlerde tutulan kadınların genel sağlığını izlemek için haftalık ağırlıklar belirlendi. Önceki çalışmamızda, anne diyet grupları arasında vücut ağırlığında herhangi bir değişiklik gözlenmemiştir4. Şekil 1 , çiftleşmeden önceki dört...

Bu protokol, anne diyetinin yavru sağlığı sonuçları üzerindeki etkisini araştırmak amacıyla hamilelikten önce dişi farelerde diyetle folik asit veya kolin alımını azaltma yöntemini açıklar.

Maternal nutrition during pregnancy and lactation plays an important role in the neurodevelopment of offspring. One-carbon (1C) metabolism, which centers ...

modifying-levels-maternal-dietary-folic-acid-or-choline-to-study

Research

JoVE Journal

Medicine

Neuroscience

Modifying Levels of Maternal Dietary Folic Acid or Choline to Study the Impact of Deficiencies on Offspring Health Outcomes

Maternal nutrition during pregnancy and lactation plays an important role in the neurodevelopment of offspring. One-carbon (1C) metabolism, which centers around folic acid and choline, as well as other B vitamins, plays a key role during the closure of the neural tube of the developing fetus. However, the impact of these maternal nutritional deficiencies during pregnancy on offspring health outcomes after birth remains relatively undefined. Furthermore, maternal dietary deficiencies in folic acid or choline may impact other health outcomes in offspring - making this a valuable model. This protocol aims to outline the procedure for inducing a deficiency in 1C metabolism in female mice through dietary modifications. Females are placed on diets at weaning, up to 2 months of age, for 4-6 weeks prior to mating and remain on diet throughout pregnancy and lactation. Offspring from these females can be evaluated for health outcomes. Females can be used multiple times to generate offspring, and tissues from females can be collected to measure for 1C metabolite measurements. This protocol provides an overview of how to induce maternal dietary deficiencies in folic acid or choline to study offspring health outcomes.

This protocol describes the method for reducing dietary intake of folic acid or choline in female mice prior to pregnancy with the objective of investigating the impact of maternal diet on offspring health outcomes.

Tıp

Inducing Maternal Dietary Deficiencies in Mice to Study the Impact on Offspring Health

To begin, obtain folic acid or choline-deficient feed from commercial rodent food manufacturers. The special diet contains 0.3 milligrams per kilogram of folic acid, or 300 milligrams per kilogram of choline by tartrate. Maintain a female mouse on this special diet for four to six weeks.Later, when the offspring are weaned from the mother, Place them on a folic acid or choline-deficient diet as well as a control diet. After euthanizing the offspring, collect the blood sample and liver tissue for biochemical analysis. Add the blood to an EDTA-coated tube and centrifuge the tube to isolate the blood plasma for determining the concentration of one-carbon metabolites.Then, transfer the cut liver tissue samples into a cryo-storage tube for snap freezing. The baseline and weekly weights of female mice on control, choline, or folic acid-deficient diets during the four weeks before mating showed no significant difference. The maternal one-carbon metabolite concentrations were slightly low in the plasma of mice subjected to folic acid-deficient diet as compared to the control mice.