Substance P-, F8Famide-, and A18Famide-like immunoreactivity in the nervus terminalis and retina of the goldfish Carassius auratus.

We re-investigated the occurrence of substance P-like immunoreactivity in the retina of the goldfish Carassius auratus using antisera to substance P and other tachykinins. Most antisera labelled a previously described single class of mono-stratified amacrine cells arborizing in layer 3 of the inner plexiform layer. Preabsorption experiments showed that these amacrine cells contained at least one tachykinin-like peptide. One antiserum (INC 353) to substance P labelled not only these amacrine cells but also fibres in layer 1 of the inner plexiform layer and fibres in the optic nerve. These fibres were identified as retinopetal projections of the nervus terminalis, in part because of colocalized labelling with antisera against gonadotropin-releasing hormone and FMRFamide. Preabsorption experiments showed that the substance P-immunoreactive material in the nervus terminalis was not substance P or any other typical tachykinin. Labelling of the nervus terminalis with INC 353 was blocked by preabsorption with two bovine FMRF-amide-like peptides, F8Famide and A18Famide, which contain a substance P(4-7)-like region. Antisera to F8Famide and A18Famide strongly labelled ganglia of the nervus terminalis and retinopetal fibres. We suggest that labelling of the nervus terminalis by antisera to substance P and FMRFamide occurs because of homologies between these antigens and a non-tachykinin, endogenous peptide that is similar to F8Famide and A18Famide.

Thyroid hormone resistance in hibernating ground squirrels, Spermophilus richardsoni. I. Increased binding of triiodo-L-thyronine and L-thyroxine by serum proteins.

Thyroid function was examined during the annual cycle of Richardson's ground squirrel, Spermophilus richardsoni. A number of facets were measured to facilitate comparison of thyroid function in active, dormant, and aroused animals. This report discusses changes in the serum thyroid hormone and binding as assessed by radioimmunoassay, equilibrium dialysis, and competitive binding assays. During the hibernation phase (both dormant and aroused), total serum T3 (trioodo-L-thyronine) and T4 (L-thyroxine) are elevated over active levels, two- to fivefold and four- to sixfold, respectively. However, in dormant squirrels, both free T3 and free T4 are reduced compared with both active and aroused phases of the annual cycle, while in aroused squirrels there is an increase in free T3 but no change in free T4 compared with active squirrels. The difference between changes in total and free thyroid hormone levels in the three groups is due to changes in serum binding of thyroid hormone. There is a more than twofold increase in the capacity of a saturable T3-binding site in serum of both dormant and aroused squirrels, and there is an increase in serum binding affinity at the low core temperature of dormant squirrels (6 degrees). Therefore, even though serum total T3 and T4 are elevated during dormancy, free T3 and T4 levels are reduced to half of the levels in active squirrels as a consequence of increased serum binding capacity and affinity. In aroused animals, however, increased serum binding capacity only partially buffers the increase in total T3 and T4, so that free thyroid hormone levels exceed those of active squirrels.

Thyroid hormone resistance in hibernating ground squirrels, Spermophilus richardsoni. II. Reduction of hepatic nuclear receptors.

Hepatic nuclear triiodo-L-thyronine (T3) receptors were studied to investigate the mechanism of thyroid hormone resistance in Richardson's ground squirrels, Spermophilus richardsoni, during the hibernation phase of the annual cycle. The cycle is divided into an active phase and a hibernation phase, the latter composed of alternating dormancy and arousal bouts. In addition to animals in these three states, a group of cold-exposed animals was also examined (those animals held at 6 degrees which showed no indications of entering hibernation). Binding of T3, to squirrel hepatic nuclei from all groups, was characterized as high affinity, Kd ranging from 111 to 267 pM, and low capacity, 50 to 314 fmol T3/mg DNA. Based on these data, other criteria examined, and models established in the literature for other species, this binding site has been tentatively identified as a T3 receptor. Receptor concentrations in nuclei from dormant and aroused squirrels were only 15-20% of the concentration in active animals. There were no differences in the affinity of the T3 receptor over the annual cycle estimated by the in vitro assay at 24-26 degrees. The reduction in nuclear T3 receptors, together with the previously reported increase in serum binding of thyroid hormone, provides an explanation for thyroid hormone resistance during the hibernation phase in S. richardsoni.