scholarly journals Immunocytochemical localization of luteinizing hormone-releasing hormone (LHRH) and LHRH-like material in brain. Influence of fixation solution at various pH values.

1983 ◽  
Vol 31 (11) ◽  
pp. 1329-1332 ◽  
Author(s):  
D T Piekut
Keyword(s):  
Luteinizing Hormone ◽  
Pineal Gland ◽  
Ph Value ◽  
Acidic Ph ◽  
Immunocytochemical Localization ◽  
Luteinizing Hormone Releasing Hormone ◽  
Releasing Hormone ◽  
Ph Values ◽  
Rat Pineal Gland ◽  
The Brain

The amount and distribution of luteinizing hormone-releasing hormone (LHRH) immunoreactive (ir) material in the hypothalamus, and of a LHRH-like immunostained substance in the rat pineal gland were examined employing Bouin's and Zamboni's fixation solutions at various pH values, and antisera generated against synthetic LHRH. The hypothalamic LHRH immunoreaction product is optimally visualized in this study when the pH value of the fixation solution is basic; conversely, the pineal LHRH-like ir material is seen only with an acidic pH value of the fixation solution. The present study demonstrates that the pH value of the fixation solution contributes substantially to the amount and distribution of some peptides visualized immunocytochemically in the brain.

scholarly journals Immunocytochemical analysis of the rat pineal gland using antisera generated against analogs of luteinizing hormone-releasing hormone (LHRH).

1982 ◽  
Vol 30 (2) ◽  
pp. 106-110 ◽  
Author(s):  
D T Piekut ◽  
K M Knigge
Keyword(s):  
Luteinizing Hormone ◽  
Pineal Gland ◽  
Median Eminence ◽  
Luteinizing Hormone Releasing Hormone ◽  
Immunocytochemical Analysis ◽  
Releasing Hormone ◽  
Immunoreactive Material ◽  
Lhrh Agonists ◽  
C Terminus ◽  
Rat Pineal Gland

Antisera generated against synthetic luteinizing hormone-releasing hormone (LHRH) or analogs of LHRH, agonists and antagonist, were used to stain the rat pineal gland and the median eminence of the hypothalamus. Of the LHRH analogs used, two revealed immunoreactive material in median eminence and not pineal, two stained pineal only, and three stained median eminence and pineal. Our observations suggest that the immunoreactive material in the pineal gland may be an LHRH-like substance whose affinity characteristics are different from that of hypothalamic LHRH. The affinity properties of hypothalamic LHRH for antibody appear to reside in the C-terminus of the molecule, whereas binding of the antigen in the pineal to antibody appear to be influenced by an N-terminal modification of the LHRH molecule.

scholarly journals Immunocytochemical analysis of the rat pineal gland using antisera generated against luteinizing hormone-releasing hormone (LHRH).

1981 ◽  
Vol 29 (5) ◽  
pp. 616-622 ◽  
Author(s):  
D T Piekut ◽  
K M Knigge
Keyword(s):  
Luteinizing Hormone ◽  
Pineal Gland ◽  
Median Eminence ◽  
Luteinizing Hormone Releasing Hormone ◽  
Immunocytochemical Analysis ◽  
Releasing Hormone ◽  
Immunoreactive Material ◽  
Rat Pineal Gland

Seven different antisera generated against synthetic luteinizing hormone-releasing hormone (LHRH) were used to stain pineal gland and median eminence of the hypothalamus of normal rats. The LHRH immunogen complexes used to generate these antisera produced either N- or C-terminal directed antibodies or antisera that recognized both portions of the hormone molecule. Of these, four revealed LHRH-like immunoreactive material in the median eminence and pineal gland, two stained median eminence and not pineal, and one was positive in pineal only. These observations suggest that immunoreactive material in pineal gland may be an LHRH-like substance whose affinity characteristics are different from that of hypothalamic LHRH.

scholarly journals Immunocytochemical localization of luteinizing hormone-releasing hormone (LHRH) in Vibratome-sectioned brain.

1981 ◽  
Vol 29 (2) ◽  
pp. 247-254 ◽  
Author(s):  
S A Joseph ◽  
D T Piekut ◽  
K M Knigge
Keyword(s):  
Luteinizing Hormone ◽  
Immunocytochemical Localization ◽  
Fiber Distribution ◽  
Fixed Tissue ◽  
Luteinizing Hormone Releasing Hormone ◽  
Final Reaction Product ◽  
Releasing Hormone ◽  
Tissue Sections ◽  
Dehydration Processes ◽  
Sectioned Tissue

Immunocytochemical localization of neuropeptides such as luteinizing hormone-releasing hormone (LHRH) is generally performed on Bouin's fixed tissue sections, following tissue dehydration in alcohols and embedment in paraffin. When the final reaction product accurately reflects content and distribution of the neuropeptide has not been examined carefully. Our data indicate a decrease in radioimmunoassayable LHRH content of brain fixed in Bouin's solution and a further significant reduction following dehydration with alcohol. In order to circumvent this loss of hormone, sections of fixed brain were cut on a Vibratome at 30 micron and collected in phosphate-saline buffer. These Vibratome sections revealed a significantly greater amount of overall immunoreactivity and fiber distribution as compared to paraffin-embedded sectioned tissue. These results suggest that certain neuropeptides, soluble in alcohol, can be extracted during dehydration processes necessary for paraffin embedment.

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