Combination of immunohistochemical and in situ hybridization methods to reveal tyrosine hydroxylase and oxytocin and vasopressin mRNAs in magnocellular neurons of obese Zucker rats.
Co-localization of chemical messengers in the same neuron is linked to neurochemical plasticity and has been studied extensively [B. Meister, M.J. Villar, S. Ceccatelli, T. Hökfelt, Localization of chemical messengers in magnocellular neurons of the hypothalamic supraoptic and paraventricular nuclei: an immunohistochemical study using experimental manipulation, Neuroscience 37 (1990) 603-633; B. Meister, R. Cortés, M.J. Villar, M. Schalling, T. Hökfelt, Peptides and transmitter enzymes in hypothalamic magnocellular neurons after administration of hyperosmotic stimuli: comparison between messenger RNA and peptide/protein levels, Cell Tissue Res. 260 (1990) 279-297]. Obese Zucker rats display an example of such a phenomenon expressing an enzyme of catecholamine synthesis-tyrosine hydroxylase (TH)-in magnocellular neurons (MCN) of supraoptic and paraventricular nuclei of hypothalamus [S. Fetissov, F. Marsais, S. Nicolaïdis, A. Calas, Expression of tyrosine hydroxylase in magnocellular hypothalamic neurons of obese (fa/fa) and lean heterozygous (Fa/fa) Zucker rats, Mol. Brain Res. 50 (1997) 314-318]. To understand the biological role of TH in MCN of obese Zucker rat, we studied TH expression in relation to the vasopressinergic and oxytocinergic neurons. We present a protocol of double labelling including immunohistochemical for TH and in situ hybridization for OT and VP mRNA. The described protocol can be applied for detection of co-localized expressions of a broad range of chemical brain messengers and proteins.
Fetissov
S O
SO
Department of Surgery, SUNY Health Science Center, Syracuse 13210, USA. fetissos@mailbox.hscsyr.edu
Marsais
F
F
D000818:Animals; D007031:Hypothalamus / Q000378:metabolism* / Q000473:pathology; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D009474:Neurons / Q000378:metabolism; D009765:Obesity / Q000235:genetics / Q000378:metabolism*; D010121:Oxytocin / Q000235:genetics*; D010904:Pituitary Gland, Posterior / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D011924:Rats, Zucker / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics*; D014667:Vasopressins / Q000235:genetics*
D012333:RNA, Messenger; D014667:Vasopressins; D010121:Oxytocin; D014446:Tyrosine 3-Monooxygenase