Abstract
Successful culturing of neurons from adult animals has been historically difficult for a relatively long time. In this study, we reported the development of a novel method for the isolation and the culture of major pelvic ganglion (MPG) neurons from adult rat. The cultured cells were identified by neuron morphology and staining with neuronal marker (neurofilament-200, NF-200). The results demonstrate that the new protocol we used was reliable in obtaining a relatively high yield of MPG neurons. Furthermore, it improves the speed and simplicity in neuronal isolation. The viability of neurons can be maintained for about 2 weeks, which should be sufficient for investigating physiological and pathological processes occurring in mature major pelvic ganglia. And this may provide a useful assessment to currently available techniques for the culture of adult neurons.
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References
Andrews TJ, Thrasivoulou C, Nesbit W, Cowen T (1996) Target-specific differences in the dendritic morphology and neuropeptide content of neurons in the rat SCG during development and aging. J Comp Neurol 368:33–44
Brewer GJ (1997) Isolation and culture of adult rat hippocampal neurons. J Neurosci Methods 71:143–155
Brewer GJ, Espinosa J, McIlhaney MP, Pencek TP, Kesslak JP, Cotman C, Viel J, McManus DC (2001) Culture and regeneration of human neurons after brain surgery. J Neurosci Methods 107:15–23
Cowen T, Jenner C, Xiao Song Gu, Budi Santoso AW, Gavazzi I (1997) Responses of mature and aged sympathetic neurons to laminin and NGF: an in vitro study. Neurochem Res 22:1003–1011
Dail WG, Evan AP, Eason HR (1975) The major ganglion in the pelvic plexus of the male rat: a histochemical and ultrastructural study. Cell Tissue Res 159:49–62
Greenwood D, Coggeshall RE, Hulsebosch CE (1985) Sexual dimorphism in the numbers of neurons in the pelvic ganglia of adult rats. Brain Res 340:160–162
Henderson CE (1996) Role of neurotrophic factors in neuronal development. Curr Opin Neurobiol 6:64–70
Keast JR (1995) Visualization and immunohistochemical characterization of sympathetic and parasympathetic neurons in the male rat major pelvic ganglion. Neuroscience 66:655–662
Keast JR (1999) Unusual autonomic neurons: chemistry, connections and plasticity of the pelvic ganglia. Int Rev Cytol 193:1–69
Keast JR (2006) Plasticity of pelvic autonomic ganglia and urogenital innervation. Int Rev Cytol 248:141–208
Keast JR, de Groat WC (1989) Immunohistochemical characterization of pelvic neurons which project to the bladder, colon, or penis in rats. J Comp Neurol 288:387–400
Keast JR, Booth AM, de Groat WC (1989) Distribution of neurons in the major pelvic ganglion of the rat which supply the bladder, colon or penis. Cell Tissue Res 256:105–112
Nangle MR, Keast JR (2011) Semaphorin 3A inhibits growth of adult sympathetic and parasympathetic neurones via distinct cyclic nucleotide signalling pathways. Br J Pharmacol 162:1083–1095
Perry SW, Norman JP, Litzburg A, Gelbard HA (2004) Antioxidants are required during the early critical period, but not later, for neuronal survival. Neurosci Res 78:485–492
Ren D, Miller JD (2003) Primary cell culture of suprachiasmatic nucleus. Brain Res Bull 61:547–553
Steers WD, Ciambotti J, Erdman S, de Groat WC (1990) Morphological plasticity in efferent pathways to the urinary bladder of the rat following urethral obstruction. J Neurosci 10:1943–1951
Steers WD, Kolbeck S, Creedon D, Tuttle JB (1991) Nerve Growth factor in the urinary bladder of the adult regulates neuronal form and function. J Clin Invest 88:1709–1715
Tuttle JB, Steers WD (1992) Nerve growth factor responsiveness of cultured major pelvic ganglion neurons from the adult rat. Brain Res 588:29–40
Uchida Y, Tomonaga M (1985) Effects of nerve growth factor and heart cell conditioned medium on neurite regeneration of aged sympathetic neurones in culture. Brain Res 348:100–106
Ullian EM, Sapperstein SK, Christopherson KS, Barres BA (2001) Control of synapse number by glia. Science 291:657–661
Wanigasekara Y, Keast JR (2005) Neurturin has multiple neurotrophic effects on adult rat sacral parasympathetic ganglion neurons. Eur J Neurosci 22:595–604
Wanigasekara Y, Kepper ME, Keast JR (2003) Immunohistochemical characterisation of pelvic autonomic ganglia in male mice. Cell Tissue Res 311:175–185
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This study is supported by National Basic Research Program of China (2003CB515304).
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Cheng, S., Yang, X., Zhang, Y. et al. Culture of major pelvic ganglion neurons from adult rat. Cytotechnology 65, 663–669 (2013). https://doi.org/10.1007/s10616-012-9515-5
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DOI: https://doi.org/10.1007/s10616-012-9515-5