Twelve biotinylated lectins and an avidin-biotin-peroxidase method were used to detect and localize specific carbohydrate residues on formalin-fixed, paraffin-embedded female canine mammary gland sections. Histologic sections from 3 lactating and 7 nonlactating mixed-breed dogs (age 5.6 +/- 0.35 years) were incubated with Arachis hypogea agglutinin (peanut agglutinin; PNA), Concanavalia ensiformis agglutinin (conA), Dolichos biflorus agglutinin (DBA), Glycine max agglutinin (SBA), Griffonia simplicifolia agglutinin-I (GS-I), Lens culinaris agglutinin (LCA), Lycopersicon esculentum agglutinin (LEA), Phytolacca americana mitogen (pokeweed mitogen; PWM), Ricinus communis agglutinin-I and-II (RCA-I and -II), Triticum vulgaris (WGA), and Ulex europaeus agglutinin-I (UEA-I). Each lectin had a specific binding pattern, except SBA and DBA. In nonlactating glands, PNA, conA, LEA, and UEA-I stained duct cells in a linear-binding pattern, with a mean percentage of positive ducts per section of 28.7 (+/- 0.6), 65.7 (+/- 0.3), 100 (+/- 0), and 8.4 (+/- 0.2), respectively. Strong apical, lateral, basal, and cytoplasmic positivity on duct cells was seen after incubation of the sections with RCA-I, RCA-II, and WGA in all ducts. In acinar cells, the binding pattern and the staining distribution of all the lectins studied were similar to those in duct cells. However, for PNA, conA, and UEA-I, the mean percentage of positive lobules per section was 33.7 (+/- 0.9), 62 (+/- 0.5), and 10.5 (+/- 0.2), respectively. In glands from lactating dogs, conA and UEA-I did not stain. The cytoplasm of all myoepithelial cells was moderately stained with RCA-I, RCA-II, and WGA. Endothelial cells stained with GS-I, PWM, RCA-I, RCA-II, WGA, conA, and LCA. The extracellular matrix, especially the periacinar and periduct regions, and the interstitial fibroblasts were positive for LCA, RCA-I, RCA-II, and WGA. Peripheral unmyelinated nerve fibers of the nipple were strongly positive for GS-I, PWM, RCA-1, RCA-II, and WGA. Some of the lectins used (ie, PNA, conA, UEA-I, GS-I, PWM, and LEA) appear to have selective staining of mammary gland structures that seems to be correlated with various physiologic functions. The contrasting binding pattern of lectins specific for the same sugar indicates a lack of knowledge of interactions between lectins and carbohydrate residues in tissue sections.

The compound nerve action potential (CNAP) of the superficial peroneal nerve of dogs was investigated to determine: (1) the influence of the stimulation technique on the configuration of the CNAP, with particular attention to late components; (2) the fiber diameter (FD) distribution; and (3) the relationship between FD distribution and CNAP configuration, by reconstruction of CNAP made on the basis of FD distributions. The CNAP were evoked in 9 dogs under halothane anesthesia by 2 stimulation methods: percutaneous needle electrode stimulation and direct stimulation of the exposed superficial peroneal nerve. Recordings were made with percutaneous needle electrodes. Full nerve cross sections of 7 superficial peroneal nerves were prepared for FD morphometric analysis. Reconstruction of CNAP were made on the basis of the FD distributions. Late components of the CNAP could be evoked with either stimulation method, but only with a stimulus intensity of 3 to 5 times maximal for the main (early) component of the CNAP. The FD histograms of 7 analyzed nerves had bimodal distribution. In 5 nerves, peaks were at 4.2 to 4.5 micrometer and 9.0 to 10.0 micrometer with 60% of the fibers in the small-diameter group. In 2 nerves with lower maximal conduction velocities, peaks were shifted toward smaller values. The CNAP reconstructions made by use of FD data closely resembled actual recordings when a fifth-order polynomial function was applied to the relationship between nerve conduction velocity and FD. Reconstructions made by use of 1 or 2 linear functions did not accurately resemble actual recordings. The results indicate clinical sensory electroneurographic recordings can provide accurate information regarding both large- and small-diameter fibers, if adequate stimulus intensities are used. To understand the recorded potential more completely, further studies are needed to determine the effects of volume conduction on configuration of the CNAP. It should then be possible to estimate FD distributions even more accurately by analyzing CNAP of normal nerves, or of diseased nerves in which the normal relation between FD and conduction velocity is preserved.