Neuropeptide gene expression in hypothalamic magnocellular neurons of normal and hypophysectomized rats: a combined immunohistochemical and in situ hybridization study.
Hypothalamic magnocellular neurons of the paraventricular and supraoptic nuclei contain several peptides and non-peptide putative neurotransmitters co-existing with vasopressin and oxytocin. However, the functional role of these substances is still unknown. In the present paper the temporal course of changes in the expression of vasopressin, oxytocin, galanin, cholecystokinin, dynorphin and tyrosine hydroxylase in magnocellular hypothalamic neurons of rats subjected to hypophysectomy was examined. Following different survival times the animals were processed either for immunohistochemistry with antibodies against the above mentioned peptides or for in situ hybridization with synthetic oligonucleotide probes complementary to the mRNAs encoding for the peptides. The results obtained showed a marked rise in vasopressin mRNA levels at two days followed by a decrease up to 36 days of survival. Oxytocin mRNA responded to the lesion with a transient decrease, with its lowest values between five and seven days. This was followed by a recovery which almost reached normal values at 36 days of survival. The results also showed a marked, transient activation of the synthetic pathway for galanin and cholecystokinin. The numbers of cells expressing these peptides were maximal between five and seven days, and the respective mRNA levels were significantly increased at these survival times. This was followed by a decrease in the amount of galanin- and cholecystokinin-like immunoreactivity as well as in the levels of their respective mRNAs. Dynorphin-like immunoreactivity showed a course similar to that of galanin and cholecystokinin in operated animals. However, the amounts of dynorphin mRNA were significantly increased at two days, but were followed by a reduction at five days and remained low throughout the different survival times tested. The experiments performed with the tyrosine hydroxylase antibodies and probe showed undetectable levels of the enzyme and its mRNA in normal and hypophysectomized animals. These results demonstrate that, in magnocellular hypothalamic neurons, expression of several peptides occur in differential ways after hypophysectomy. The possibility is discussed that these changes represent part of the mechanisms underlying the process of degeneration and regeneration known to occur in magnocellular hypothalamic neurons after hypophysectomy.
Villar
M J
MJ
Department of Histology and Neurobiology, Karolinska Institutet, Stockholm, Sweden.
Meister
B
B
Cortés
R
R
Schalling
M
M
Morris
M
M
Hökfelt
T
T
D000818:Animals; D002766:Cholecystokinin / Q000378:metabolism; D004399:Dynorphins / Q000378:metabolism; D019004:Galanin; D015870:Gene Expression; D007016:Hypophysectomy; D007031:Hypothalamus / Q000166:cytology / Q000378:metabolism*; D007032:Hypothalamus, Anterior / Q000166:cytology / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D009474:Neurons / Q000378:metabolism*; D009479:Neuropeptides / Q000096:biosynthesis / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010121:Oxytocin / Q000378:metabolism; D010286:Paraventricular Hypothalamic Nucleus / Q000166:cytology / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D014667:Vasopressins / Q000378:metabolism
D009479:Neuropeptides; D010455:Peptides; D012333:RNA, Messenger; D014667:Vasopressins; D010121:Oxytocin; D004399:Dynorphins; D019004:Galanin; D002766:Cholecystokinin