Tight junction composition is altered in the epithelium of polycystic kidneys.

Kidney cysts in autosomal dominant polycystic kidney disease (ADPKD) undergo progressive enlargement together with luminal fluid secretion. This involves active, uphill transcellular Cl(-) transport which drives passive Na(+) and water secretion. Implicit in this mechanism is the assumption that the paracellular permeability of the cyst epithelium to Cl(-) must be very low. Claudins are tight junction (TJ) transmembrane proteins that determine the ion selectivity of paracellular barriers. The aim of this study was to determine the expression and localization of claudins within renal cysts in a mouse hypomorphic model of ADPKD and in human patients. We found that the majority of cysts were of collecting duct origin. Claudins normally expressed in collecting duct (3, 4, 7, 8, and 10) were found in small cysts. However, only claudin-7 persisted at substantive levels in the dedifferentiated epithelium of large, presumably late-stage cysts, where it was localized both at the TJ and basolaterally. The constitutively expressed TJ proteins, ZO-1 and occludin, were also abundantly expressed and correctly localized, suggesting that the basic infrastructure of the TJ is preserved. A previous study suggested that claudin-7 may function as a paracellular Cl(-) barrier. We postulate that the role of claudin-7 in ADPKD is to seal the paracellular route in Cl(-)-secreting cyst epithelium, preventing backleak of Cl(-), and that it thereby plays a permissive role in fluid secretion and cyst growth.

Dissociaton of conditioned eye and limb responses in the cerebellar interpositus.

Thompson and colleagues have demonstrated that the lateral interpositus nucleus of the cerebellum is the essential locus for the classical conditioning of the somatic eyeblink response. Preliminary studies reported that lesioning the cerebellar interpositus nucleus ipsilateral to the side of training also appears to abolish conditioned limb flexion responses. Previous studies have suggested that the interpositus nucleus is somatotopically organized with the eye being represented laterally and the hindlimb medially. Presently, we employed a double dissociation paradigm to examine the effects of muscimol (a GABA(A) agonist) injections on eyeblink versus limb flexion conditioned responses in the ipsilateral cerebellar interpositus nucleus of New Zealand white rabbits. For eyeblink conditioning, the conditioned stimulus (CS) was a 14-V lamp bulb and the unconditioned stimulus (US) was a 3-psi corneal airpuff to the left eye. For limb flexion conditioning, the CS was a 1-kHz, 85-95 dB SPL tone and the US was a 3- to 5-mA shock to the upper left hindlimb. Upon training on both responses to a 60-100% criterion, the rabbits were then tested on eyeblink and limb flexion responses after injections of muscimol (0.1-0.3 mul of a 0.01- to 1.0-M solution) into either the lateral (eyeblink) or medial (limb flexion) interpositus nucleus. We have been able to successfully decrease or abolish the percent conditioned responses (CRs) of both the eyeblink and limb flexion conditioning selectively without affecting the other. These results thus lend further support for the notion of the existence of a somatotopic map in the interpositus nucleus for learning.

Transfer of learning but not memory after unilateral cerebellar lesion in rabbits.

Unilateral lesion of the cerebellum in rabbits completely and permanently abolishes previous learning and prevents new learning ipsilateral to the lesion. However, when training continues on the contralateral side, there is substantial savings in that it takes few trials to learn. This observation may imply that the memory survives the lesion. Rabbits were classically conditioned for an eyelid response and then the ipsilateral interpositus nucleus of the cerebellum was lesioned. Then the rabbits were trained on the contralateral side. There is no savings on the contralateral side without first trying to train on the lesioned side. The authors conclude that the survival of a memory after the lesion probably does not account for the rapid transfer, but rather that the act of trying to train on the lesioned side in previous studies first induces a new memory on the contralateral side.

Lesions of the cerebellar interpositus nucleus abolish both nictitating membrane and eyelid EMG conditioned responses.

Both nictitating membrane extension and eyelid EMG activity were measured during classical conditioning of rabbits to tone-airpuff pairings. Both measures were highly correlated. Over trials, learning criterion was met earlier with eyelid EMG activity than with nictitating membrane extension. Within a trial, eyelid EMG preceded and was more robust than nictitating membrane extension. The rabbits were lesioned in the cerebellar interpositus nucleus and then trained for up to 26 days. Detailed analyses of tone alone trials demonstrate that the lesion abolished conditioned responses for both measures. These data confirm that conditioned responses are abolished by lesion of the cerebellar interpositus nucleus.