<meta charset="utf8"></head><body>山羊空肠倒陷囊中钠依赖性组氨酸吸收

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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+uptake+of+amino+acids+by+the+intestine.">The uptake of amino acids by the intestine.</a> BBA - Biochim Biophys Acta. 14 (1), 80-84 (1954).</li><li>Pauly, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=&#220;ber+die+Konstitution+des+Histidins:+I.+Mitteilung.">&#220;ber die Konstitution des Histidins: I. Mitteilung.</a> Biological Chemistry. 42 (5-6), 508-518 (1904).</li><li>Wilson, T. H., Wiseman, G. <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+sacs+of+everted+small+intestine+for+the+study+of+the+transference+of+substances+from+the+mucosal+to+the+serosal+surface.">The use of sacs of everted small intestine for the study of the transference of substances from the mucosal to the serosal surface.</a> J Physiol. 123 (1), 116-125 (1954).</li><li>Barthe, L., Woodley, J. F., Kenworthy, S., Houin, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+everted+gut+sac+as+a+simple+and+accurate+technique+to+measure+paracellular+transport+across+the+small+intestine.">An improved everted gut sac as a simple and accurate technique to measure paracellular transport across the small intestine.</a> Eur J Drug Metab Pharmacokinet. 23 (2), 313-323 (1998).</li><li>Alam, M. 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<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1.5 mL Microcentrifuge Tubes</td>
			<td>TARSONS</td>
			<td>500020</td>
			<td></td>
		</tr>
		<tr>
			<td>10 mL Test Tubes</td>
			<td>BOROSIL</td>
			<td>9800U04</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL Sterile Falcon Tubes</td>
			<td>TARSONS</td>
			<td>546041</td>
			<td></td>
		</tr>
		<tr>
			<td>500 mL Beaker</td>
			<td>BOROSIL</td>
			<td>10044977</td>
			<td></td>
		</tr>
		<tr>
			<td>500 mL Conical Flask</td>
			<td>BOROSIL</td>
			<td>691467</td>
			<td></td>
		</tr>
		<tr>
			<td>D-Glucose</td>
			<td>SRL</td>
			<td>42738</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital Spectrophotometer</td>
			<td>SYSTRONICS</td>
			<td>2710</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>EMSURE</td>
			<td>1009831000</td>
			<td></td>
		</tr>
		<tr>
			<td>Finpipettes</td>
			<td>THERMOFISHER</td>
			<td>4642090</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass Stirrer Rod</td>
			<td>BOROSIL</td>
			<td>9850107</td>
			<td></td>
		</tr>
		<tr>
			<td>L-Histidine&#160;</td>
			<td>SRL</td>
			<td>17849</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>SRL</td>
			<td>41721</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrile Gloves</td>
			<td>KIMTECH</td>
			<td>112-4847</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri Dish&#160;</td>
			<td>TARSONS</td>
			<td>460090</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline (ph 7.4)</td>
			<td>SRL</td>
			<td>95131</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette Tips</td>
			<td>ABDOS</td>
			<td>P10102</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Carbonate</td>
			<td>SRL</td>
			<td>89382</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Nitrate&#160;</td>
			<td>SRL</td>
			<td>44618</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Phosphate Dibasic (anhydrous)</td>
			<td>SRL</td>
			<td>53046</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Phosphate Monobasic (anhydrous)</td>
			<td>SRL</td>
			<td>22249</td>
			<td></td>
		</tr>
		<tr>
			<td>Sulphanilic Acid&#160;</td>
			<td>SRL</td>
			<td>15354</td>
			<td></td>
		</tr>
	</tbody>
</table>

demonstration of membrane transport of histidine using goat intestinal inverted sacs: an experiential pedagogical tool for undergraduates

Biological cells are surrounded by a membrane lipid bilayer that separates the intracellular cytosol from the extracellular content. The membrane serves as a semipermeable barrier that regulates the movement of solutes1. Transport across biological membranes is affected by a solute&#39;s permeability coefficient, which depends on several factors, including the concentration and charge of the solute. In general, solutes move through the membrane using three mechanisms (Figure 1): Passive diffusion, facilitated diffusion, and active transport2. Simple diffusion is the process by which soluble, uncharged, and non-polar solutes pass down their concentration gradient through a semipermeable membrane (Figure 1A). Membrane proteins do not assist in this process since it involves the movement of solutes from a region of higher concentration to a region of lower concentration. The rate of diffusion is based on Fick&#39;s Law3. On the other hand, facilitated diffusion is a protein-dependent transport wherein the membrane allows only selective solutes to pass along a concentration gradient without the expenditure of energy (Figure 1B). This kind of transport is specific and differs from simple diffusion in exhibiting saturation kinetics.
Active transport is a protein-dependent transport of molecules against their concentration gradient, i.e., from the region of lower concentration to a region of higher concentration with the use of ATP or ion gradients (Figure 1C). When the transporter hydrolyses ATP, the transport is termed primary active transport (Figure 1C; left panel). Another form of active transport is secondary active transport (Figure 1C; right panel). In secondary active transport, solutes are moved based on an electrochemical gradient. It takes place when a transporter protein couples the movement of an ion (typically Na+) down its concentration gradient with the movement of another molecule or an ion against its concentration gradient. This kind of solute movement can be a co-transport (Symport) wherein both the solute and the ion move in the same direction or an exchange (Antiport), in which case the solute and the ion move in opposite directions.
The dietary amino acids and monosaccharides from food sources are absorbed in the small intestine. The small intestine can be functionally divided into three segments: Duodenum, Jejunum, and Ileum (Figure 2). Absorption of solute occurs throughout the small intestine, with maximum absorption occurring at the jejunum and at the proximal end of the ileum. The intestinal enterocytes are polarized cells, and the tight junctions connecting two adjacent cells create two distinct membrane sites - the basolateral and the apical membrane site (Figure 2). The absorption of luminal solutes generated by digestion occurs on the apical membrane site4.
Histidine transport in the intestinal enterocytes at the apical membrane is an example of a secondary active symport that is sodium-dependent. At the basolateral end, the histidine entering the enterocyte moves down the concentration gradient into the hepatic portal circulation. The intracellular concentrations of sodium within the enterocyte are maintained at 12 mmoles/L1, which is lower than the extracellular/luminal concentrations, due to the active pumping of sodium out of the cell by Na+ K+ATPase located on the basolateral membrane (Figure 3). At the apical membrane of enterocytes, the B0AT and the sodium neutral amino acid transporter (SNAT) 5 are the major transporters that not only transport histidine but also amino acids such as asparagine and glutamine in a sodium-dependent cotransport5,6. Another transport protein called Large Amino acid Transporter (LAT)1 present at the basolateral membrane of enterocytes transports large neutral amino acids such as leucine, tryptophan, tyrosine, and phenylalanine across the membrane7.
With the objective of teaching the concept of membrane transport integrated with techniques such as spectrophotometry and routine biochemical assays through experiential learning pedagogy, it is imperative to develop methodologies that can not only demonstrate the concept in easy-to-understand terms but also enable participatory learning for undergraduate students. Currently, there are limited resources available to the students for hands-on engagement to learn such concepts of biochemistry. Here, we report a simple protocol for demonstrating Histidine transport across goat intestinal membrane that is easy to reproduce in undergraduate laboratories and can be adapted for evaluating the transport of other metabolites as well. More importantly, the method utilizes inexpensive materials in an undergraduate laboratory, thereby enabling experiential learning in even the simplest of laboratory settings.

<meta charset="utf8"></head><body>作者聚焦：使用离体组氨酸摄取教授主动运输的体验工具

使用山羊肠倒囊演示组氨酸的膜转运：本科生的体验式教学工具

Here we propose a pedagogical tool for undergraduate students of biology where we are demonstrating the concept of membrane transport. What we are actually doing here is the, with the objective of showing the transport of histidine, which is an amino acid across the enterocyte membrane, in a simple, easy to perform, inexpensive experimental setting, which can be done in almost any laboratory. The current challenge that we face in an undergraduate laboratory is the ban that has been imposed on animal dissections.Now because of this, what we are doing here is procuring the intestines from a common vendor and intestines that we do get, however, have a problem because they are not from syngeneic animals, and therefore there would be a variability in the animals samples that we get. So currently there are no simple and easy to perform experiments that can facilitate learning about the concepts of memory and transport. Most of the methods available use hazardous chemicals that are not feasible for teaching learning in a UG setting.So this technique demonstrates an X vivo setup that doesn't rely on cell-free extracts, but a tissue sample which can be derived from different available mammalian sources. So that's the obvious advantage to our method.

Histidine is an essential amino acid that is also a precursor for metabolites implicated in the immune system, pulmonary ventilation, and vascular ...

demonstration-membrane-transport-histidine-using-goat-intestinal

Research

JoVE Journal

Biochemistry

Demonstration of Membrane Transport of Histidine using Goat Intestinal Inverted Sacs: An Experiential Pedagogical Tool for Undergraduates

812 次观看.  University of Delhi. 在这里，我们为生物学本科生提出了一种教学工具，我们在其中演示了膜传输的概念。我们在这里实际做的是，目的是在一个简单、易于执行、廉价的实验环境中展示组氨酸（一种氨基酸）穿过肠上皮细胞膜的运输，这几乎可以在任何实验室中完成。我们目前在本科实验室面临的挑战是禁止动物解剖。正因为如此，我们在这里所做的是从共同的...