Mycobacterium marinum causes long-term subclinical granulomatous infection in immunocompetent leopard frogs (Rana pipiens). These granulomas, organized collections of activated macrophages, share many morphological features with persistent human tuberculous infection. We examined organs of frogs with chronic M. marinum infection using transmission electron microscopy in conjunction with immunohistochemistry and acid phosphatase cytochemistry to better define the bacterium-host interplay during persistent infection. Bacteria were always found within macrophage phagosomes. These phagosomes were often fused to lysosomes, in sharp contrast to those formed during in vitro infection of J774 macrophage-like cells by M. marinum. The infected macrophages in frog granulomas showed various levels of activation, as evidenced by morphological changes, including epithelioid transformation, recent phagocytic events, phagolysosomal fusion, and disintegration of bacteria. Our results demonstrate that even long-term granulomas are dynamic environments with regard to the level of host cell activation and bacterial turnover and suggest a continuum between constantly replicating bacteria and phagocytic killing that maintains relatively constant bacterial numbers despite an established immune response. Infection with a mutant bacterial strain with a reduced capacity for intracellular replication shifted the balance, leading to a greatly reduced bacterial burden and inflammatory foci that differed from typical granulomas.

Two-dimensional crystals of cholera toxin bound to receptors in a lipid membrane give diffraction extending to 15 A resolution. Three-dimensional structure determination reveals a ring of five B subunits on the membrane surface, with one-third of the A subunit occupying the center of the ring. The remaining mass of the A subunit appears to penetrate the hydrophobic interior of the membrane. Cleavage of a disulfide bond in the A subunit, which activates the toxin, causes a major conformational change, with the A subunit mostly exiting from the B ring.

ncui MID/OR MTCRCSTMG © The Mi era be am Analysis Society (MAS) Council recently established a special fund, the Fiori fund, to support a touring speaker program to honor the memory of Dr, Chuck Fiori, whose untimely passing was felt by many microscopists. The touring speaker program is available to Local Affiliate Societies and will specifically address the needs of technicians and other related topics. The Council intends that this program be permanent and will support it with MAS Council funds. Contributions to the Fiori fund are welcome at any time and should be sent to the MAS Treasurer,

Extensive t i g h t j u n c t i o n s a r e l o c a t e d between overlaps o f plasma membranes o f c a p i l l a r y e n d o t h e l i a l c e l l s . M i c r o p i n o c y t o t i c v e s i c l e s a r e few i n number and a r e seen as i n v a g i n a t i o n s on t h e A f a c e and as p r o t r u s i o n s on t h e B face. appearance o f these c e l l s i s c o n s i s t e n t w i t h t h e i r t h i n s e c t i o n appearance and t h e i r r o l e as t h e s i t e o f t h e b l o o d -b r a i n b a r r i e r .The f r e e z e -f r a c t u r e The c a p i l l a r y endothelium i s b e l i e v e d t o r e p r e s e n t t h e s i t e o f t h e b l o o d -b r a i n b a r r i e r i n t h e c e r e b r a l c o r t e x o f a d u l t mammals (Reese and Karnovsky, '67; Brightman and Reese, ' 6 9 ) . I n t h e present i n v e s t i g a t i o n , t h e appearance o f f r e e z e -f r a c t u r e and f r e e z e -e t c h r e p l i c a s o f c a p i l l a r y was f r a c t u r e d i n a B a l z e r ' s BA360M u n i t (Moor e t a l . , '61; Branton, '66) a t -115O C o r f r a c t u r e d and etched a t -looo C. endothelium o f t h e c e r e b r a l c o r t e x i n mice has been c o r r e l a t e d w i t h t h e u l t r a s t r u c t u r e seen i n t h i n s e c t i o n s and w i t h t h e f r e e z e -f r a c t u r e appearance o f o c c l u d i n g j u n c t i o n s i n o t h e r t i s s u e s . MATERIALS AND METHODSwere cleaned i n Purex and dimethylformamide, and r i n s e d f i r s t i n H2SO4 and One-mm cubes o f t i s s u e were immersed i n 20% g l y c e r o l o v e r n i g h t p r i o r

The development of synapses in the visual cortex (VC) and superior colliculus (SC) of the rabbit has been examined with the electron microscope. In both areas, the number of synapses reaches adult levels by 20--25 days of postnatal age, but the development in the visual cortex is delayed in comparison to that in the superior colliculus. When S synapses (spheroidal vesicles, asymmetric thickening) are compared with F synapses (flattened vesicles, symmetric thickening), even greater differences are seen. In both the VC and SC, S synapses develop earlier than F synapses, though there is considerable overlap. Of interest is that fact that synapses in the visual cortex seem to overshoot their adult levels late in development, suggesting that an excess of synapses may be formed in this system. Multiple synapses, probably of retinal origin, increase in the first 3 weeks of synaptic development in the SC, but never are present in significant proportions in the VC. Synapse formation most often is characterized by formation of a junction and a postsynaptic thickening, followed by acquisition of synaptic vesicles. After 15 days, there is only a small number of such "non-vesicle synapses" in either the SC or VC.