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In Vivo Microinjection and Electroporation of Mouse Testis
Published on: August 23, 2014
Nerve growth factor mRNA and protein in the testis and epididymis of mouse and rat
C Ayer-LeLievre1, L Olson, T Ebendal
1Department of Medical Genetics, Uppsala University, Sweden.
Abstract:
In situ hybridization using beta-nerve growth factor (NGF) DNA probes was used to demonstrate NGF mRNA in spermatocytes and early spermatids of adult mouse. NGF mRNA-containing cells were also identified in the epithelium of convoluted ducts in mouse corpus epididymidis. Blot-hybridization analysis of RNA prepared from mouse testis and epididymis as well as from rat epididymis confirmed the presence of a 1.3-kilobase (kb) NGF mRNA in these tissues. In the rat testis, however, only a 1.5-kb NGF mRNA was found, corresponding in size to a minor NGF mRNA detected in the rat brain, heart, and epididymis. By using affinity-purified anti-NGF antibodies, NGF-like immunoreactivity was observed in germ cells of rat and mouse testis and in the lumen of epididymis. Extracts of both mouse epididymis and testis stimulated fiber outgrowth in cultured sympathetic ganglia, and the effect was blocked by antibodies to mouse NGF. A two-site enzyme immunoassay showed the presence of 10 and 70 ng of NGF per g of tissue in the mouse testis and epididymis, respectively. Furthermore, RNA blot analysis showed the presence of mRNA for the NGF receptor in mouse testis. These results suggest a nonneurotrophic role for NGF in the male reproductive system, possibly in survival maturation and/or motility of spermatozoa.
Insights
Nerve growth factor (NGF) mRNA and protein are present in the male reproductive system. This suggests a non-neuronal role for NGF in sperm maturation and motility.
Area of Science:
- Reproductive Biology
- Neuroscience
- Molecular Biology
Background:
- Nerve growth factor (NGF) is crucial for neuronal development and survival.
- The presence and function of NGF in the male reproductive system are not well understood.
Purpose of the Study:
- To investigate the presence and localization of NGF and its receptor in the male reproductive system.
- To explore the potential role of NGF in male reproductive functions.
Main Methods:
- In situ hybridization to detect NGF mRNA in mouse reproductive tissues.
- Blot-hybridization to analyze NGF mRNA size.
- Immunohistochemistry using anti-NGF antibodies.
- Bioassays to assess the biological activity of tissue extracts.
- Enzyme immunoassay to quantify NGF levels.
- RNA blot analysis for NGF receptor mRNA.
Main Results:
- NGF mRNA and NGF-like immunoreactivity were found in mouse and rat germ cells and epididymal epithelium.
- A 1.3-kb NGF mRNA was detected in mouse testis and epididymis, while a 1.5-kb form was found in rat testis.
- Tissue extracts stimulated nerve fiber outgrowth, which was blocked by anti-NGF antibodies.
- NGF levels were quantified in mouse testis and epididymis.
- mRNA for the NGF receptor was present in mouse testis.
Conclusions:
- NGF is present in the male reproductive system, suggesting a non-neurotrophic role.
- NGF may be involved in sperm survival, maturation, and/or motility.
